大鼠脾脏来源内皮祖细胞对C6胶质瘤细胞增殖能力影响的体外研究.docVIP

大鼠脾脏来源内皮祖细胞对C6胶质瘤细胞增殖能力影响的体外研究 方靖琴，张伟国，马长锁 （400042 重庆，第三军医大学大坪医院野战外科研究所放射科） [摘要] 目的 研究体外内皮祖细胞对C6胶质瘤细胞增殖能力的影响。方法 取SD大鼠脾脏采用密度梯度离心法获取内皮祖细胞，通过分化过程中的形态特征、内皮祖细胞吞噬乙酰化低密度酯蛋白和结合荆豆凝集素-1的特异性和免疫荧光检测细胞表面抗原CD34和CD31的表达进行鉴定。将内皮祖细胞条件培养基加至C6胶质瘤细胞的常规培养基中观测其对C6胶质瘤细胞增殖的影响；分别采用内皮祖细胞条件培养基和常规培养基孵育C6胶质瘤细胞24h，流式细胞仪检测2组的细胞周期。结果 激光共聚焦鉴定DiI-acLDL和FITC-UEA-1双染色阳性细胞即为正在分化的内皮祖细胞；免疫荧光检测内皮祖细胞一致性表达CD34和CD31。随着内皮祖细胞条件培养基含量的增加，2个检测时间点各组的吸光值也随着增加，且50%内皮祖细胞条件培养基组D（490）值(2.018±0.220)、(2.388±0.448)均高于对照组(1.163±0.103)、(1.106±0.174)，且差别有统计学意义(P＜0.01)；50%含有内皮祖细胞条件培养基培养的C6胶质瘤细胞的增殖率即S+G2/M明显高于对照组 (P＜0.05)。结论 体外内皮祖细胞条件培养基能促进C6胶质瘤细胞的增殖，其机制可能与其分泌的各种血管生成类细胞因子有关。 [关键词] 内皮祖细胞；C6胶质瘤细胞；增殖；体外 [中图法分类号] [文献标志码] A Effect of endothelial progenitor cells on the proliferation of C6 glioma cells under co-culture system in vitro Fang Jingqin, Zhang Weiguo, Ma Changsuo (Department of Radiology, Institute of Surgery Research , Daping Hospital,Third Military Medical Universaty, Chongqing,400042,China [Abstract] Objective Study the effect of endothelial progenitor cells on the growth of C6 glioma cells in vitro. Methods The endothelial progenitor cells(EPCs)were obtained from the spin of healthy SD rats with combined density gradient centrifugalization with adherence screening method. The EPCs were characterization through morphologic characteristics, specific for DiI-acLDL uptaking and Lectin binding and the expression of CD34,CD31 detected by immunofluorescence. Combine the endothelial progenitor cells conditional medium with convention medium to assess its effect on the proliferation of C6 glioma cells After cultured 24h by the endothelial progenitor cells conditional medium and conventional medium respectively, flow cytometry detected the cell cycle. Results After 10 days cultured, the attached cells were able to present “line-like” structure, the adherent cells were double positive for DiI-acLDL uptaking and FITC-UEA-1 binding by direct flourescent staining unde